3-keto-delta4-6-cyanoamino steroids



"The 17-position may bear the position: I

United States Patent Q r 3,201,426 S-KETQ-M-fi-(EYANQAMINO STEROIDS George A. Boswell, J22, Newark, and Richard M. Scribner, Wilmington, Del., assignors to E. I. du Pont de Nernours audfiompany, Wilmington, Del., a corporation of Delaware No Drawing. Filed Aug. 27, 1963, Ser. No. 304,946

8 Claims. (Cl. 260-397.2)

A -6-cyanoamino atomic grouping,

These products are preparedby'a process which coniprises oxidizing under essentially anhydrous, alkaline conditions, a 3-hydroxy-5,6-N-cyanoaziridino steroid, Le, a S-hydroxy steroid in which the steroid nucleus is fused at the 5 and 6 carbon atoms to an aziridine nucleus whose nitrogen atom has its third valence satisfied by a cyano group. This'reaction, which is a combination of oxidation and ring-opening reactions, is representedby the following equation,'in which only the, portion of the steroid comprising positions 3 to disshown:

In this discussion, the spatial configuration (0: or B) of the cyanoamino group and its precursor, the N-cyanoaziridino group, isomitted. The configuration" has not been proved rigorously. The initial 3 -hydroxy group can have either the a or e configuration.

The 3-ketoA -6-cyanoamino steroids of this inventio may bear substituents only in the 10, 13 or 17 positions. The l3-position is substituted with a methyl group, and the IO-position may bear a hydrogen or a methyl group. following. groups in the 3,291,426 Patented Aug. 17, 1965 ice groups without affecting acid-sensitive substituents present in the steroid molecule. Inthe present-case, however, it was unexpectedly found that, under these conditions, opening of the aziridine ring-with formation of a 6-cyanoamino group and of a double bondbetween the 4 and 5 carbon atoms occurs simulttneously with the oxidation of the 3-hydroxyl group. 7

The oxidizing agent of choice to effect this combined reaction is a chromium trioxide /tertiary nitrogen base complex of the type previously described in the literature, for example, by Sisler et al., J. Am. Chem. Soc., 70, 3827 (1948), and in British Patent 733,808. Suitable examples are the CrO, complexes of pyridine, the picolines, the lutidines, quinoline, diethylformamide and triethylamine. Of these, the preferred reagent is the CrO /pyridine complex, which has the composition C1O -2C H N, and, for simplification, the process will be discussed with reference to this specific reagent in the 'following'paragraphs; A

reaction with the N-cyanoaziridino steroid, although an additional inert solvent can be used if desired.

The oxidation reaction does not require elevated temperatures and it is most conveniently conducted in the temperature range of 0-30 0, although moderate heat- -ing-,'e.g., up to 75 C.,, can be applied if desired. Substantial conversions. are obtained after a few hours con tact at. room temperature, and it is usually unnecessary to. prolong the reaction time beyond 24 hours. Other secondary hydroxyl groups that may be present in the steroid molecule besides the 3-hydroxy group are also usually converted to carbonyl groups during the reaction. The reaction product is most conveniently isolated by pouring the reaction mixture into acidified water or, preferably, into water buttered to a slightly acid pH, followed by extraction of both of the resulting aqueous and solid phases with an appropriate organic solvent. The extract is evaporated to yield the 3-keto-A -6-cyanoamino Y steroid, which can be purified by recrystallization if while the at position can be --H, CH or -OAc. 'OAc represents an acetoxy group. In addition, the 17 position may be substituted with a carbonyl (:0) group.

7 The compounds of the invention are prepared by. oxidizing a 3-hydroxy-5,6-N-cyanoaziridino steroid, having the above substituents. 1

an alkaline liquid medium. Such conditions have previously been used in the steroid art to oxidiie hydroxyl 1962, which consists in reacting cyanogen azide with a B-hydroxy steroid having an intracyclic double bond between the 5 and 6 carbons, in accordance with the illustrative simplified equation solution of the unsaturated 3-hydroxy steroid in an inert solvent such as acetonitrile, ethyl acetate, toluene, methylene chloride, carbon tetrachloride, etc., with a solution to warm slowly to room temperature.

of cyanogen azide in an inert solvent such as one of those already mentioned. The cyanogen azide is used in amounts of at least one-half mole per mole of unsaturated steroid, normally in at least molar equivalent, and preferably in slight to moderate excess thereover. The reaction can be conducted at temperatures ranging from about 25 C. to about 75 C., but temperatures between 40 and 60 C. are preferably used. Its progress can be followed by determining the amount of nitrogen. evolved. The reaction product can be isolated and purified by chromatography on neutral activated alumina and/or crystallization from an appropriate solvent.

The cyanogen azide used to prepare the 3-hydroxy- 5,6-N-cyanoaziridino steroids is prepared by reacting a caynogen halide, preferably cyanogen chloride, with a metal, ammonium or organoarnmonium azide, preferably sodium azide, under anhydrous conditions, at -a temperature in the range of 50 C. to +25 C. The cyanogen azide product is a colorless, unstable liquid which, when isolated, is explosively sensitive to heat or shock. However, its solutions in organic solvents are not explosive and can be stored, handled and used with safety. A

typical preparation of cyanogen azide in solution is as follows: a

A mixture of sodium azide (16.25 g., 0.25 mole) and dry acetonitrile (48.5 g.) is placed in a flask equipped with a condenser, thermometer, magnetic stirrer, gas inlet tube and nitrogen bubbler. The flask is cooled to 10 to C. and ice-water or a coolant at 5 to 0 C. is circulated through the condenser. Cyanogen chloride (24.0 g., 0.39 mole) is distilled into the flask with good stirring at such a rate as to maintain the temperature below 12 C. After completion of the addition, the mixture is stirred for 0.5 hour at 0l0 C. and then allowed Excess cyanogen chloride is removed by evacuation through the cooled condenserat 95-120 mm. pressure for about 1 hour, with stirring. The solution which remains in the flask is filtered under nitrogen and the filter cake is washed twice 'with a total of ml. of dry acetonitrile. The combined filtrate is a solution of cyanogen azide in acetonitrile which can be used directly or stored under nitrogen at C. for subsequent use.

Solutions of cyanogen azide in any other anhydrous solvent such as ethyl acetate, benzene, acetone, etc., can. be prepared in a similar manner.

The following examples illustrate the invention.

Example 1.-6-cyan0amino-4-pregnene-3,20-dione (6 -cyan0amin0pr0gester0ne) The starting material used in this example was prepared as follows:

To a solution of 8.0 g. (25.3 millimoles) of 3-hydr-oxy- S-pregnene-ZO-one (pregnenolone) in 100 ml. of ethyl acetate was added 10 ml. (28.5 millimoles) of 2.85 M cyanogen azide in ethyl acetate. The reaction mixture was heated at 52i2 C. for 46 hours, during which time 660 ml. (27 millimoles) of gas was evolved. Fltration of the reaction mixture and evaporation of the filtrate gave 9.2 g. of amber gum. This was dissolved in 13 ml. of ethyl acetate and the solution was absorbed on 200 g. of neutral alumina (activity 111). Elution with petroleum ether, then with benzcne/ petroleum ether mixtures first gave about 2.3 g. (29%) of unchanged pregnenolone. Continued elution with 25/75 and 50/50 diethyl ether/benzene mixtures gave 3.0 g. of crude 3-hydroxy- 5,6-N-cyanoaziridinopregnane-ZO-one, crystallization of which from acetone-hexane gave 1.68 g. (24% yield) of pure product, M.P. 16l.5-162.5 C., [e1 ,+46 (acetone, c.=l.l5).

Analysis.-Calcd for C H N O C, 74.12; H, 9.05; N, 7.86. Found: C, 74.46; H, 9.25; N, 7.68.

The infrared spectrum agreed with the assigned structure, showing absorptions at 3400 cm. (OH), 2200 cm.- (N-CEN), and 1700 cm? C 0).

A solution of 2.5 g. of this steroid in 20 ml. of pyridine was added to the complex prepared from 2.5 g. of chromium trioxide and 30 m1. of pyridine. After stirring for 6 hours at room temperature, the brown reaction mixture was poured into 350 ml. of Water containing g. of sodium dihydrogen phosphate (pl-I 5). The aqueous mixture was filtered through sintered glass to give a brown solid and an aqueous filtrate. The filtrate was extracted with three ml. portions of ethyl acetate, and the filter cake was extracted with three 150 ml. portions of boiling ethyl acetate. The combined extracts were washed with 10% aqueous sodium dihydrogen phos- Ultraviolet: x523? 233 m (e=12,900)

Example 2.-2l-acetoxy-6-cyanoamin0-4-pregnene- The starting material used in this example was prepared as follows:

T 0 a solution of 9.35 g. (25 millimoles) of 21-acetoxy- 3-hydroxy-5-pregnene-20-one in ml. of ethyl acetate was added 12 ml. (34 millimoles) of 2.85 M cyanogen azide in ethyl acetate. The reaction mixture was heated at 50-53 C. for 44 hours, during which time 870 ml. (36 millimoles) of gas was evolved. The mixture was filtered to separate some solid material and the filter cake was washed with hot ethyl acetate. The combined ethyl acetate filtrates were evaporated under reduced pressure to give 12.1 g. of gummy residue which was redissolved in 25 ml. of ethyl acetate. This solution was absorbed on 400 g. of neutral alumina (activity IV).

Elution with petroleum ether, benzene, and finally 1:1

ethyl acetate-benzene gave, in the latter eluate, a colorless glass that, on crystallization from acetone-ether, afforded 2.87 g. (28% yield) of crystalline 21-acetoxy- 5,6 N-cyanoaziridino-3-hydroxypregnane-20-one, M.P.

V cm.- (NCEN); 1750 cm? and 1730 cm? (:0).

A mixture of 1.2 g. of this steroid with thecomplex prepared from 8 g. of chromium trioxide and 22 ml. of pyridine was stirred at room temperature for 22 hours,

after which the reaction product was isolated by the procedure described in Example 1. There was obtained 0.80 g. (65% yield) of 21-acetoxy-6-cyanoamino-4-pregnone-3,20-dione (also called 6-cyanoamino desoxycorticosterone acetate), M1. 153-156 C. after crystallization from acetone-benzene.

Analysis.Calcd for C I-1 N 0 C, 69.88; H, 7.82; N, 6.79. Found: C, 71.43; H, 7.74; N, 6.31.

Infrared: 3190 crn.- (NH); 2210 cm." (CEN);

1745, 1720 and 1675 cm. (ester, ketone and conjugated i C 0); 1610 cm.- (@C); and 1230 cm? (ester (#0).

Ultraviolet: x512? 232 111;; (e 12,100) Example 3.6-cyan0amin0-17,6-hydroxy-17a-methyl-4- andr0stene-3-0ne 6-cyan0amino-17a-mezhyltestosterone I l jlfi (G i l used in this example was preandrostene-3fl,l7fi=diol ih-175 ml. of ethyl acetate and 15.0 ml. (42 millimoles) of a 2.85 M solution of cyanogen azide in ethyl acetate was heated at i2 C. for 48 hours, during which time la total of 965 ml. of gas (95% of theory) was evolved. The reaction mixture was filtered and the filter cake} was washed with warm ethyl acetate. The combined i iltrate and washings were evaporated, leaving about 10 g. of a light yellow solid which was redissolved in 25 mllof ethyl acetate. This solution was absorbed on 600 g. of neutral alumina (activity IV).

Elution with benzene and then with 1:5 ethyl acetate benzene gave first 0.96 g.? of unchanged starting material, then a glass-like solid which was crystallized from ethanol by slow addition of n-hexane-to give 4.11 g. (36% yield) tals melting at 185-186 C.

glassy residue of 6-cyanoamino-l7 8-hydroxy-I'M-methyl- 4-androstene-3-one. After three crystallizations from ethanol-water, the product was obtained as colorless crys- It was soluble in 5% aqueous sodium hydroxide, insoluble in aqueous sodium carbonate." It contained one-half mole of ethanol of crystallization.

Analysis.Calcd for C H N O -1/2C H OH: C, 72.4; H, 9.10; N, 7.67. Found: C, 72.42; H, 8.82; N, 7.68.

Infrared: 3400 cm.- (OH); 3180 cmr (NH); 2200 -cm. -(NCEN) 1665 CH1.1 (conjugated C D); and 1610 cm. (C=C).

Ultraviolet: in? 232 m $13,700

Example 4.-6-cyan0amin0-4-ch0lestene-3-0ne \AZY h 1 15 I OSQI ll ON ON The starting material used in this example was prepared as follows:

A solution of cholesterol (19.3 g., 50 millimoles) in 135 ml. of ethyl acetate was dried over anhydrous calcium sulfate and the solution was filtered. To the filtrate was added 25 ml. (70 millirnoles) of 2.85 M cyanogen azide in ethyl acetate and the mixture was heated at 50 53 C. for 50 hours, during which time a total of 1730 ml. (70 millimoles) of gas was evolved. Filtration of the reaction mixture and evaporation of the'filtrate to dryness gave a gummy residue whichwas redissolved in 65 m1. of benzene. The solution was absorbed on 600 g. of neutral alumina (activity III). Elution with benzene, then with 30:70 ether-benzene gave 1.6 g. of oily product consisting chiefly of unchanged cholesterol. 'Continued elution with ether-benzene (50:50 by volume) gave an oil that, on crystallization from cyclohexane-petroleum ether, afforded 4.1 g. (20% yield) of 5,6-N-cyanoaziridinocholestane-3fi-ol as feathery crystals, M.P. 142.5- 7

' isolated by the procedure described in the preceding exof 5,6-N-cyanoaziridino-17ot-methylandrostane 3,8,175-

diol, M.P. 108112 C., --29 (acetone, c.=2.3). This product contained one moleof ethanol of crystallization.

Analysis.Calcd for C H N O -C H OH: C, 70.73; H, 9.81; N, 7.17. Found: C, ,70.55;H, 9.72; N, 7.03.

The infrared spectrum supportedthe assigned structure, exhibiting a band at 2200 cm. (NCEN).

A solution of 0.70 g. of the above steroid in 8 ml. of pyridine was added to the complex prepared from 0.70 g. of chromium trioxide and 7 ml. of pyridine. The reaction mixture was stirred for 18 hours at room temperature and then poured into a solution of 10 ml. of concentrated sulfuric acid in 120 ml. of water. The resulting aqueous and solid phases were each extracted with two 75 ml. portions of ethyl acetate. The combined extracts were evaporated, giving 0.328 g. (47% yield) of a colorless amples except that the precipitating liquid was water acidified to a pH of 1 with hydrochloric acid. There was obtained 0.329 g. of a white solid. This solid was dissolved in cyclohexane and petroleum ether was added.

On standing overnight an oily solid precipitated. This was separated by decantation and redissolved in 3 ml. of cyclohexane. Chromatography of this solution on 15 g.

of neutral alumina (activity 111) gave, on elution with 1:1 ether-benzene, 0.192 g. of 6-cyanoamino-4-cholestene- 3-one, M.P. 97.598.5 C. after recrystallization from cyclohexane-petroleum ether.

Analysis.-Calcd for C H N O: N, 6.60. Found: N,

The infrared spectrum supported the assigned structure, showing absorptions at 3200 cm? (NH); 2200 cm? (CEN); 1670 cm. (conjugated C=O); and 1610 cm." (C=C).

Ultraviolet}: 705.53? 233 m (e=12,200)

steroids which can be prepared (with or without additional, conventional transformations such as, e.g., esterification, etherification or acetalization of hydroxyl groups) bythe described procedure, such as that illustrated in Example I, include those listed below opposite the formula of the starting 3-hydroxy-5,6-N-cyanoaziridin0 steroid.

Starting material Reaction product i CN OAc

17B-Ac0toxy-6-cyanoamino-4- audrostene-B-one.

I A CH3 6-cyauoamiuo-l-stigmnsteuc- I me.

E O f C N 0 I] Q s-cyanoamino-4-androstcne- 3, 17-dione.

fi-cyanoamino.17B-hydr0xyl7a-methyl-4-estrene-3-one.

$35 $113 /CH3 (|)H(CH2)2CHCH 6Cyanoamiuo-l-ergostenc-E- one.

Starting material Reaction product CH3 JH Ac ---o. .c

' 17a,20a-Diaeetoxy6-cyanoamino-4-pregnene-3-one.

17a-Acetoxy-6-cyanoamino4- pregnene-3,20-dione.

N AN

Attachment of the cyanoamino function at the 6-position of the steroid nucleus, a key position especially in the biologically most active 3-keto-A system, affects the biological properties of the steroid molecule. For instance, introduction of the 6-cyanoamino group results in the elimination or reduction of the primary hormone effects such as androgenic activity. Thus, whereas the 6- unsubstituted hormonal steroids such as progesterone or 17 a-methyltestosterone show progestational or androgenic activity, respectively, the 6-cyanoamino-3-keto-A steroids of this invention possess antiandrogenic activity. This was indicated by tests on male rats which showed a involution of the ventral prostate gland following subcutaneous administration of the G-cyanoaminoprogesterone of Example .1 and of the 6-cyanoamino-l7a-methyltestosterine of Example 3, While the testes and levator ani muscles remained essentially unchanged. In addition, administration of 6-cyanoaminoprogesterone to estrogenprimed immature female rabbits followed by histological examination of their uteri showed that this compound had little or no progestati-onal activity. This specificity shows usefulness of these compounds in the chemical treatment of prostatic hypertrophy, and it has the advantage of permitting administration of these products in relatively massive doses.

As many apparently widely diiferent embodiments of this invention may be made without departing from the spirit and scope thereof, it is to be understood that this invention is not limited to the specific embodiments thereof except as defined in the appended claims.

The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows:

1. A steroid of the formula wherein R is selected from the class consisting of methyl and hydrogen; R is selected from the class consisting of and -CH(OAc)CH R" is selected from the class consisting of H, --CH and 0A0, wherein Ac is acetoxy; and R and R taken together represent =0.

2. 6-cyanoamino-4-pregnene-3,20-dione.

3. 21-acetoxy-6-cyanoamino-4-pregnene-3,20-dione.

4. 6-cyanoaminod7fi-hydroxy 17a methyl 4 and-rostene-3-one.

5. 6-cyanoamino-4-cholestene-3-one.

6. A process for preparing the steroids of claim 1 which comprises oxidizing a 3-hydroxy-5,6-N-cyanoaziridino steroid having the substituents defined in claim 1, with a chromium trioxide/tertiary nitrogen base complex oxidizing agent, at a temperature of from 0 C. to 75 C.

7. The process of claim 6 in which the oxidizing agent is a chromium trioxide/pyridine complex.

8. A process for preparing 6-cyanoamino-4-pregnane- 3,20 dione which comprises reacting 3-hydroxy-5,6-N- cyanoaziridinopregnane-ZO-one with a chromium trioxide/ pyridine complex.

References Cited by the Applicant Bowers et al., J. Am. Chem. Soc., 81, 5233 (1959).

LEWIS GOTTS, Primary Examiner.

UNITED STATES PATENT OFFICE CERTIFICATE OF CORRECTION 'atent No. 3,201,426 August 17, 1965 George A. Boswell, Jr. et a1.

It is hereby certified that error appears in the above numbered pat- :nt requiring correction and that the said Letters Patent should read as :orrected below.

Column 1, line 69, for "C=N" read CN column 4, .ines 43 to 56, the formulas should appear as shown below .nstead of as in the patent:

. O\C/JH OAC /CHZOAC CrO -pyr. HO M O N blIH Ch CN olumn 5, lines 26 and 27 heading to Example 3 should appear n italics as shown below instead of as in the patent:

6cyanoamino-l76-hydroxy-17a-methyl-4-androstene3-one 6-cyanoamino-l7d-methyltestosterone olumn 5, line 44, for "l7B=diol" read l7B-diol column 6,

ine 8, for "C H N O read C I I N O same column 6,

lines 16 to 28, the right-rand formula should appear as shown below instead of as in the patent:

columns 7 and 8, in the table, first column, second formula thereof, the upper right-hand portion, for

H3 (lz n i5 CZHS cn {xi-CK 3 read H- (CH2) Z-CH-C( same table, first column, fourth formula thereof, the formula should appear as shown below instead of as in the patent:

same table, first column, fifth formula should appear as shown below instead of as in the patent:

H H i 3 3 i H3 H- (CH2)2-CH-CH column 9, line 46, for "testosterine" read testosterone column 10, lines 66 and 67 for "-pregnane-3,2O dione" read pregnene3,20dione Signed and sealed this 22nd day of March 1966.

(SEAL) Attest:

ERNEST W. SWIDER EDWARD J. BRENNER Attesting Officer Commissioner of Patents 

1. A STEROID OF THE FORMULA 